Examining post-transcriptional regulation of the segmentation clock by microRNAs

检查 microRNA 对分段时钟的转录后调节

• 批准号: 7773417
• 负责人: SUSAN E COLE
• 金额: $ 7.52万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7773417
• 关键词: 3' Untranslated Regions; Binding; Biological Assay; Cell Culture Techniques; Congenital Abnormality; Coupled; Data; Defect; Disease; Dysostoses; Embryo; Embryonic Development; Exhibits; FOLH1 gene; Feedback; Future; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; Genetic Translation; Half-Life; Hour; In Situ Hybridization; Link; Messenger RNA; Methods; MicroRNAs; Mission; Mus; Mutation; National Institute of Child Health and Human Development; Nucleotides; Pathway interactions; Pattern; Play; Post-Transcriptional Regulation; Process; Production; Public Health; RNA; RNA Binding; RNA Stability; Regulation; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; Segmentation Clock Pathway; Signal Pathway; Skeletal Development; Skeleton; Somites; Techniques; Testing; Time; Transcript; Transcriptional Regulation; Untranslated Regions; Vertebral column; base; candidate validation; falls; interest; mRNA Expression; mRNA Stability; member; notch protein; programs; research study; rib bone structure; somitogenesis

DESCRIPTION (Provided by Applicant): The vertebrate embryonic axis consists of serially repeated lements, the most evident of which are the somites. The proper production of somites during somitogenesis is critical to the formation of the axial skeleton. The process of somitogenesis is regulated in part by a genetic clock. Key genes in the Notch, Wnt, and Fgf pathways exhibit cyclic expression with a period that matches the rate of somite formation. Mutations that perturb the proper oscillation of these genes can result in congenital defects of the axial skeleton, including spondylocostal dysostoses. Thus, understanding the regulatory mechanisms that control segmentation clock function is critical to our understanding of skeletal development. Several groups have examined the transcriptional control of oscillatory gene expression linked to the segmentation clock, but comparatively little is known about the post-transcriptional mechanisms that regulate clock function. The investigator hypothesizes that microRNAs may play important roles in the post-transcriptional control of genes linked to the clock. miRNAs are small conserved RNAs that bind to target transcripts and promote mRNA turnover and/or inhibit mRNA translation. She proposes that the function of these miRNAs may contribute to the delays in the negative feedback regulatory loops that underlie oscillatory RNA expression. She has identified miRNAs that are enriched in a pattern that is consistent with potential functions in the segmentation clock, and find that many of these miRNAs are predicted to target the 3'UTRs of oscillatory genes. Identifying miRNAs that function in the segmentation clock would provide a new paradigm to understand the posttranscriptional regulation of the segmentation clock. To pursue this idea it will be critical to generate further preliminary data connecting these candidate miRNAs to potential functions in the clock. The investigator therefore proposes to examine these miRNAs during somitogenesis by validating the enrichment of these candidate miRNAs, examining their expression patterns throughout embryogenesis, and functionally testing their ability to target and regulate expression of oscillatory genes. The experiments proposed here fall directly in the scope of the R03 program, as they represent a small, self-contained project that will produce critical preliminary data for future research. Further, this research falls directly into the mission of the NICHD, examining the mechanisms by which the spatial and temporal control of gene expression during embryogenesis functions to regulate somitogenesis. It is predicted that the research proposed here will provide critical links between miRNA regulation and segmentation clock function, and will provide a base of preliminary data that will support further research into the actual functions of miRNAs during somitogenesis as well as the segmentation clock in a future R01 application. RELEVANCE: The processes of embryonic development that produce the spine and ribs are controlled by a genetic clock. When this clock is perturbed, the results can be congenital defects. This research examines different mechanisms that control the expression of genes that have been linked to this clock. By understanding how the expression of genes in the clock is controlled, we will be better able to target treatments for defects and diseases that arise from misregulation of the clock.

描述（由申请人提供）：脊椎动物胚胎轴由串行重复的雌性组成，其中最明显的是节点。 在体内生成过程中的适当产生对于轴向骨骼的形成至关重要。 生物发生的过程部分由遗传时钟调节。 Notch，Wnt和FGF途径中的关键基因表现出循环表达，其时期与体征形成的速率相匹配。 这些基因适当振荡的突变会导致轴向骨骼的先天性缺陷，包括脊柱脊髓失调。 因此，了解控制分割时钟功能的调节机制对于我们对骨骼发育的理解至关重要。 几个小组检查了与分割时钟相关的振荡基因表达的转录控制，但是对于调节时钟功能的转录后机制知之甚少。 研究者假设microRNA可能在与时钟相关的基因的转录后控制中起重要作用。 miRNA是与靶转录物结合并促进mRNA转换和/或抑制mRNA翻译的小保守RNA。 她建议这些miRNA的功能可能导致振荡性RNA表达的负反馈调节环的延迟。 她已经确定了以与分割时钟中潜在功能一致的模式富集的miRNA，并发现许多这些miRNA被预测靶向振荡基因的3'UTR。 识别分段时钟功能的miRNA将提供一个新的范式，以了解分割时钟的转录后调节。 为了提出这一想法，至关重要的是生成将这些候选miRNA与时钟潜在功能联系起来的进一步初步数据。 因此，研究者建议通过验证这些候选miRNA的富集，在整个胚胎发生过程中检查其表达模式，并在功能上测试其靶向和调节振荡基因表达的能力，从而检查这些miRNA。 这里提出的实验直接属于R03计划的范围，因为它们代表了一个小型，独立的项目，该项目将为未来的研究提供关键的初步数据。 此外，这项研究直接属于NICHD的使命，研究了胚胎发生功能中基因表达的空间和时间控制以调节体重生成的机制。 据预测，这里提出的研究将在miRNA调控和分段时钟功能之间提供关键联系，并将提供初步数据的基础，这些数据将支持对在体重生成过程中miRNA的实际功能以及未来R01应用程序中的细分时钟进行进一步研究。 相关性：产生脊柱和肋骨的胚胎发育过程由遗传时钟控制。 当这个时钟受到干扰时，结果可能是先天性缺陷。 这项研究研究了控制与该时钟相关的基因表达的不同机制。 通过了解如何控制时钟中基因的表达，我们将能够更好地靶向因时钟造成不利影响的缺陷和疾病的治疗方法。